Compounds and methods for modulating expression of SGLT2

Abstract

The present disclosure describes short antisense compounds, including such compounds comprising chemically-modified high-affinity monomers 8-16 monomers in length. Certain such short antisense compound are useful for the reduction of target nucleic acids and/or proteins in cells, tissues, and animals with increased potency and improved therapeutic index. Thus, provided herein are short antisense compounds comprising high-affinity nucleotide modifications useful for reducing a target RNA in vivo. Such short antisense compounds are effective at lower doses than previously described antisense compounds, allowing for a reduction in toxicity and cost of treatment. In addition, the described short antisense compounds have greater potential for oral dosing.

1. A compound comprising a short antisense oligonucleotide consisting of 10 to 14 linked nucleosides targeted to SGLT2, wherein said short antisense oligonucleotide comprises at least an eight consecutive nucleobase portion of the nucleobase sequence set forth in SEQ ID NO:281, and the compound is at least 95% complementary to sodium dependent glucose transporter 2 (SGLT2) encoded by SEQ ID NO: 3 as measured over the entire length of the compound. 
     
     
       2. The compound of  claim 1 , consisting of a single-stranded modified oligonucleotide. 
     
     
       3. The compound of  claim 1 , wherein the short antisense oligonucleotide is a DNA oligonucleotide. 
     
     
       4. The compound of  claim 1 , wherein said short antisense oligonucleotide is a RNA oligonucleotide. 
     
     
       5. The compound of  claim 1 , wherein said short antisense oligonucleotide consists of a nucleobase sequence of SEQ ID ON:281. 
     
     
       6. The compound of  claim 1 , wherein said short antisense oligonucleotide consists of the nucleobase sequence of SEQ ID NO:281 and comprises:
 a gap segment consisting of eight linked deoxynucleosides; 
 a 5′ wing segment consisting of two linked nucleosides; 
 a 3′ wing segment consisting of two linked nucleosides; 
 wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment, wherein each nucleoside of each wing segment comprises a 2′-O-methoxyethyl modified sugar, wherein each internucleoside linkage of said modified oligonucleotide is a phosphorothioate linkage, and wherein each cytosine of said modified oligonucleotide is a 5-methylcytosine. 
 
     
     
       7. The compound of  claim 1  wherein the short antisense oligonucleotide comprises a 2′-deoxyribonucleotide gap region flanked on each side by a wing, wherein each wing independently comprises 1 to 3 high-affinity modified nucleosides. 
     
     
       8. The compound of  claim 7 , wherein said high-affinity modified nucleosides are sugar modified nucleotides. 
     
     
       9. The compound of  claim 8 , wherein at least one of the sugar-modified nucleotides comprises a bridge between the 4′ and the 2′ position of the sugar. 
     
     
       10. The compound of  claim 8 , wherein each of said high-affinity modified nucleotides confers a ΔT m  of 1 to 4 degrees per nucleotide. 
     
     
       11. The compound of  claim 8 , wherein each of said sugar-modified nucleotides comprises a 2′-substituent group that is other than H or OH. 
     
     
       12. The compound of  claim 11 , wherein at least one of said sugar-modified nucleotides is a 4′ to 2′ bridged bicyclic nucleotide. 
     
     
       13. The compound of  claim 11 , wherein each of the 2′-substituent groups is, independently, alkoxy, substituted alkoxy, or halogen. 
     
     
       14. The compound of  claim 13 , wherein each of the 2′-substituent groups is OCH 2 CH 2 OCH 3 . 
     
     
       15. The compound  claim 9 , wherein the conformation of each of said sugar-modified nucleotides is, independently, β-D or α-L. 
     
     
       16. The compound  claim 11 , wherein each of said bridges independently comprises 1 or from 2 to 3 linked groups independently selected from -[C(R 1 )(R 2 )] n —, —C(R 1 )═C(R 2 )—, —C(R 1 )═N—, —C(═NR 1 )—, —C(═O)—, —C(═S)—, —O—, —Si(R 1 ) 2 —, —S(═O) x — and —N(R 1 )—;
 wherein x is 0, 1, or 2; 
 n is 1, 2, 3, or 4; 
 each R 1  and R 2  is, independently, H, a protecting group, hydroxyl, C 1 -C 12  alkyl, substituted C 1 -C 12  alkyl, C 2 -C 12  alkenyl, substituted C 2 -C 12  alkenyl, C 2 -C 12  alkynyl, substituted C 2 -C 12  alkynyl, C 5 -C 20  aryl, substituted C 5 -C 20  aryl, heterocycle radical, substituted heterocycle radical, heteroaryl, substituted heteroaryl, C 5 -C 7  alicyclic radical, substituted C 5 -C 7  alicyclic radical, halogen, OJ 1 , NJ 1 J 2 , SJ 1 , N 3 , COOJ 1 , acyl (C(═O)—H), substituted acyl, CN, sulfonyl (S(═O) 2 -J 1 ), or sulfoxyl (S(═O)-J 1 ); and 
 each J 1  and J 2  is, independently, H, C 1 -C 12  alkyl, substituted C 1 -C 12  alkyl, C 2 -C 12  alkenyl, substituted C 2 -C 12  alkenyl, C 2 -C 12  alkynyl, substituted C 2 -C 12  alkynyl, C 5 -C 20  aryl, substituted C 5 -C 20  aryl, acyl (C(═O)—H), substituted acyl, a heterocycle radical, a substituted heterocycle radical, C 1 -C 12  aminoalkyl, substituted C 1 -C 12  aminoalkyl or a protecting group. 
 
     
     
       17. The compound of  claim 16 , wherein each of said bridges is, independently, 4′-CH 2 -2′, 4′-(CH 2 ) 2 -2′, 4′-CH 2 —O-2′, 4′-(CH 2 ) 2 —O-2′, 4′CH 2 —O—N(R 1 )-2′ and 4′-CH 2 —N(R 1 )—O-2′- wherein each R 1  is, independently, H, a protecting group or C 1 -C 12  alkyl. 
     
     
       18. The compound of  claim 7 , wherein each of the high-affinity modified nucleosides is independently selected from bicyclic nucleotides or other 2′-modified nucleotides. 
     
     
       19. The compound of  claim 18 , wherein the 2′-modified nucleotides are selected from halogen, allyl, amino, azido, thio, O-allyl, O—C 1 -C 10  alkyl, —OCF 3 , O—(CH 2 ) 2 —O—CH 3 , 2′-O(CH 2 ) 2 SCH 3 , O—(CH 2 ) 2 —O—N(R m )(R n ) or O—CH 2 —C(═O)—N(R m )(R n ), where each R m , and R n  is, independently, H or substituted or unsubstituted C 1 -C 10  alkyl. 
     
     
       20. The compound of  claim 19 , wherein the 2′-modified nucleotide is a 2′-OCH 2 CH 2 OCH 3  nucleotide. 
     
     
       21. The compound of  claim 7 , wherein the short antisense oligonucleotide comprises at least one modified internucleoside linkage. 
     
     
       22. The compound of  claim 21 , wherein the modified internucleoside linkage is a phosphorothioate linkage. 
     
     
       23. The compound of  claim 7 , wherein each modified internucleoside linkage is a phosphorothioate internucleoside linkage. 
     
     
       24. The compound of  claim 7  that is 10-13 linked nucleosides in length. 
     
     
       25. The compound of  claim 7  that is 10-12 linked nucleosides in length. 
     
     
       26. The compound of  claim 7  that is 10-11 linked nucleosides in length. 
     
     
       27. The compound of  claim 7  that is 10 linked nucleosides in length. 
     
     
       28. The compound of  claim 7  that is 11 linked nucleosides in length. 
     
     
       29. The compound of  claim 7  that is 12 linked nucleosides in length. 
     
     
       30. The compound of  claim 7  that is 13 linked nucleosides in length. 
     
     
       31. The compound of  claim 7  that is 14 linked nucleosides in length. 
     
     
       32. The compound of  claim 7 , having a motif selected from 1-12-1; 2-10-2; 1-10-1; 1-10-2; 3-8-3; 2-8-2; 1-8-1; and 3-6-3, wherein, the first number represents the number of linked nucleosides in the 5′-wing, the second number represents the number of linked nucleosides in the gap, and the third number represents the number of linked nucleosides in the 3′ wing. 
     
     
       33. The compound of  claim 32  wherein the motif is selected from 1-10-1; 2-10-2; and 1-9-2. 
     
     
       34. The compound of  claim 7  having a motif selected from 1-1-10-2, 1-1-8-2, 1-1-6-3, and 1-2-8-2, wherein the first number represents the number of linked nucleosides in a first 5′ wing, the second number represents the number of linked nucleosides in a second 5′ wing, the third number represents the number of linked nucleosides in the gap, and the fourth number represents the number of linked nucleosides in the 3′ wing. 
     
     
       35. The compound of  claim 7  having a motif selected from 2-10-1-1, 2-8-1-1, 3-6-1-1, and 2-8-2-1, wherein the first number represents the number of linked nucleosides in the 5′ wing, the second number represents the number of linked nucleosides in the gap, the third number represents the number of linked nucleosides in a first 3′ wing, and the fourth number represents the number of linked nucleosides in a second 3′ wing. 
     
     
       36. The compound of  claim 7  having a motif selected from 1-1-8-1-1; 2-1-6-1-1; and 1-2-8-2-1, wherein the first number represents the number of linked nucleosides in a first 5′ wing, the second number represents the number of linked nucleosides in a second 5′ wing, the third number represents the number of linked nucleosides in the gap, the fourth number represents the number of linked nucleosides in a first 3′ wing and the fifth number represents the number of linked nucleosides in a second 3′ wing. 
     
     
       37. A composition comprising a compound of  claim 1  or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or diluent. 
     
     
       38. A method of treating a condition or disease associated with SGLT2 expression or overexpression in a mammal in need thereof, comprising administering to said mammal an effective amount of said short antisense oligonucleotide of  claim 1 . 
     
     
       39. The method of  claim 38 , wherein the disease is a metabolic disorder. 
     
     
       40. The method of  claim 39 , wherein the metabolic disorder is hyperglycemia, prediabetes, diabetes (type I and type II), obesity, insulin resistance, metabolic syndrome or any combination thereof. 
     
     
       41. A method of decreasing glucose levels, insulin resistance, HbA1c or any combination thereof in a human by administering the compound of  claim 1 . 
     
     
       42. A composition comprising a compound of  claim 6  or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or diluent. 
     
     
       43. A method of treating a condition or disease associated with SGLT2 expression or overexpression in a mammal in need thereof, comprising administering to said mammal an effective amount of said short antisense oligonucleotide of  claim 6 . 
     
     
       44. The method of  claim 43 , wherein the disease is a metabolic disorder. 
     
     
       45. The method of  claim 44 , wherein the metabolic disorder is hyperglycemia, prediabetes, diabetes (type I and type II), obesity, insulin resistance, metabolic syndrome or any combination thereof. 
     
     
       46. A method of decreasing glucose levels, insulin resistance, HbA1c or any combination thereof in a human by administering the compound of  claim 6 .